RESUMO
Lupus nephritis (LN) is a kidney inflammatory disease caused by systemic lupus erythematosus (SLE). NLRP3 inflammasome activation is implicated in LN pathogenesis, suggesting its potential targets for LN treatment. Melatonin, an endogenous indoleamine, is considered an important multitasking molecule that has been reported to have anti-inflammatory effects by inhibiting nuclear factor-kappa B (NF-κB)-mediated inflammatory responses in vivo. This molecule has also protective effects against the activation of the inflammasomes and, in particular, the NLRP3 inflammasome. Thus, this work evaluated the effect of melatonin on morphological alteration and NLRP3 inflammasome activation in LN pristane mouse models. To evaluate the melatonin effects in these mice, we studied the renal cytoarchitecture by means of morphological analyses and immunohistochemical expression of specific markers related to oxidative stress, inflammation and inflammasome activation. Our results showed that melatonin attenuates pristane-induced LN through restoring of morphology and attenuation of oxidative stress and inflammation through a pathway that inhibited activation of NLRP3 inflammasome signaling. Our data clearly demonstrate that melatonin has protective activity on lupus nephritis in these mice that is highly associated with its effect on enhancing the Nrf2 antioxidant signaling pathway and decreasing renal NLRP3 inflammasome activation.
Assuntos
Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Inflamassomos/efeitos dos fármacos , Nefrite Lúpica/tratamento farmacológico , Melatonina/uso terapêutico , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Feminino , Inflamassomos/metabolismo , Nefrite Lúpica/etiologia , Nefrite Lúpica/metabolismo , Melatonina/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Terpenos/toxicidadeRESUMO
Multiple Myeloma (MM) is a hematological malignancy characterized by the clonal proliferation of plasma cells within the bone marrow. Diagnosing MM presents considerable challenges, involving the identification of plasma cells in cytology examinations on hematological slides. At present, this is still a time-consuming manual task and has high labor costs. These challenges have adverse implications, which rely heavily on medical professionals' expertise and experience. To tackle these challenges, we present an investigation using Artificial Intelligence, specifically a Machine Learning analysis of hematological slides with a Deep Neural Network (DNN), to support specialists during the process of diagnosing MM. In this sense, the contribution of this study is twofold: in addition to the trained model to diagnose MM, we also make available to the community a fully-curated hematological slide dataset with thousands of images of plasma cells. Taken together, the setup we established here is a framework that researchers and hospitals with limited resources can promptly use. Our contributions provide practical results that have been directly applied in the public health system in Brazil. Given the open-source nature of the project, we anticipate it will be used and extended to diagnose other malignancies.
Assuntos
Mieloma Múltiplo , Humanos , Medula Óssea/patologia , Brasil , Hematologia/métodos , Aprendizado de Máquina , Mieloma Múltiplo/diagnóstico , Mieloma Múltiplo/patologia , Redes Neurais de Computação , Plasmócitos/patologiaRESUMO
BACKGROUND: To investigate gene expression of podocyte-specific proteins in urine of diabetes and prediabetes subjects and the association of these proteins with albuminuria. METHODS: Fifteen controls, 19 prediabetes, and 67 diabetes subjects were included. Messenger RNA of nephrin, podocin, podocalyxin, synaptopodin, TRPC6, alpha-actinin-4, and TGF-ß1 were measured using RT-PCR. Podocyte marker expression was correlated with albuminuria, glycemic control, and renal function. The diagnostic performance of the genes used to detect increased albuminuria was assessed using ROC curves and Poisson regressions. RESULTS: Podocyte marker expression was significantly higher in diabetic subjects. Urinary nephrin was correlated with increasing levels of albuminuria; risk of albuminuria increased by 20% for every one-unit increase in the log10 of nephrin mRNA. Nephrinuria was found in 53%, 71%, and 90% of normo-, micro-, and macroalbuminuric diabetes subjects, respectively (p = 0.023). Urinary nephrin, podocalyxin, TRPC6, podocin, and alpha actinin-4 were correlated with glycemic control and albuminuria but not with renal function. CONCLUSIONS: Diabetes subjects had higher urinary mRNA levels of podocyte proteins than nondiabetic subjects, even the normoalbuminuric patients. Nephrinuria was correlated with diabetic nephrophathy stage and predicted pathological albuminuria. Urinary mRNA levels of podocyte markers of prediabetic subjects did not differ from controls.
Assuntos
Glicemia/metabolismo , Diabetes Mellitus/metabolismo , Nefropatias Diabéticas/metabolismo , Podócitos/metabolismo , Proteinúria/metabolismo , Proteoma/metabolismo , RNA Mensageiro/urina , Adulto , Feminino , Teste de Tolerância a Glucose , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The prevalence of Leptospirosis in goat herds of the State of Minas Gerais has seldom been studied. The present research had as its objectives: (1) investigate the seroprevalence of Leptospirosis in the county of Uberlândia, MG, (2) verify the Leptospirosis serovars, and (3) identify the risk factors associated with infection on the farms examined. Serum samples from 230 animals in 11 properties were tested using the microscopic agglutination test. An epidemiological examination furnished data for analysis regarding the risk factors. The prevalence of Leptospirosis was found to be 31.3% with variation from 1:100 to 1:800. The most frequent serovars were: Autumnalis (30.30%), Tarassovi (19.20%), Pyrogenes (13.13%), and Icterohaemorrhagiae (11.11%). The ages and races of the animals were among the risk factors found to be significantly correlated (P<0.05) with infection. At the farm level, the intensity of production, use of salaried workers, and association of other animals were all found to be related with the frequency of Leptospirosis. The results demonstrated that inadequate management was a factor which favored the occurrence of infection in the region of the study.
Assuntos
Anticorpos Antibacterianos/sangue , Doenças das Cabras/epidemiologia , Leptospira/classificação , Leptospira/imunologia , Leptospirose/veterinária , Animais , Brasil/epidemiologia , Feminino , Doenças das Cabras/diagnóstico , Doenças das Cabras/microbiologia , Cabras , Testes de Hemaglutinação/veterinária , Leptospira/isolamento & purificação , Leptospirose/diagnóstico , Leptospirose/epidemiologia , Leptospirose/microbiologia , Masculino , Prevalência , Fatores de Risco , Estudos SoroepidemiológicosRESUMO
Corynebacterium pseudotuberculosis is a facultative intracellular pathogen that uses various mechanisms to survive within macrophages. In phagocytosis, this survival can be attributed to the ability to inhibit phagosome-lysosome fusion. In this fusion, some proteins, including Rabs GTPases, are involved in the maturation process and are responsible for regulating membrane vesicle trafficking. Thus, to better understand these mechanisms, the capacity of biofilm-producing and non biofilm-producing strains of Corynebacterium pseudotuberculosis for modulating the expression of endosomal proteins GTPases Rab 5 and Rab 7 was evaluated in an in vitro study of infection of goat macrophages. Blood was collected from ten Canindé goats, infected with biofilm-producing and non biofilm-producing strains of C. pseudotuberculosis. Blood cells were separated in colloidal silica-polyvinylpyrrolidone gradients (GE Healthcare®). These cells were maintained at 37 °C, with 5% of CO2. After differentiation, macrophages were infected with the mentioned strains. The bacterial pellets were marked with Rab 5 and Rab 7 antibodies, and their expression was observed by flow cytometry. Both strains of C. pseudotuberculosis (biofilm-producing and non biofilm-producing) were observed to be capable of altering the expression of Rab proteins in macrophages cultivated in vitro. Macrophages from the animals infected with the biofilm-producing strain had an increase in the expression of Rab 5 protein, mainly when these macrophages were treated with the non biofilm-producing strain. The same mechanism was shown to function with Rab 7 protein, however at a lower intensity of expression when compared with Rab 5.
Assuntos
Infecções por Corynebacterium , Corynebacterium pseudotuberculosis , Animais , Biofilmes , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/genética , Macrófagos , Fagocitose , Proteínas rab de Ligação ao GTP/genética , Proteínas rab de Ligação ao GTP/metabolismoRESUMO
Systemic lupus erythematosus (SLE) is an autoimmune disease that affects multiple organs; lupus nephritis (LN) is one of the most severe complications of SLE. In the kidneys, an intense inflammatory reaction affects the glomeruli and tubular interstitium. Uric acid has been considered a key molecule in the pathogenesis of some conditions such as metabolic syndrome, hypertension, and kidney disease as it is produced by injured cells and promotes immune-inflammatory responses. In this regard, high serum uric acid concentrations may be involved in the activation of some inflammatory pathways, associated with kidney damage in SLE. Therefore, the purpose of this article was to review the main physiological mechanisms and clinical data on the association between serum uric acid and kidney damage in SLE. Scientific evidence indicates that hyperuricemia has the potential to be an adjuvant in the development and progression of kidney manifestations in SLE. Uric acid may promote the activation of inflammatory pathways and the formation and deposition of autoantibodies in kidneys, leading to a reduction of glomerular filtration rate. Other potential mechanisms of this association include the presence of polymorphisms in the urate transporters, metabolic syndrome, use of some medications, and other situations associated with a reduced renal excretion of uric acid.
Assuntos
Nefropatias , Lúpus Eritematoso Sistêmico , Nefrite Lúpica , Humanos , Rim , Ácido ÚricoRESUMO
As part of a clinical practice which aims to alleviate the psychological suffering of children in situations of migration and/or uprooting, a discussion can be undertaken regarding the cultural aetiologies of migrant families whose children present autism spectrum disorders. The transcultural context enables families to travel between worlds as well as share their collective imagination regarding psychological suffering using graphic expression and narrative elements.
Assuntos
Transtorno do Espectro Autista , Migrantes , Criança , Humanos , Narração , Pais , Estresse PsicológicoRESUMO
BACKGROUND: Renal fibrosis is the result of the interaction of cellular and molecular pathways, which is induced by sustained glomerular injury and involves the podocytes and multiple profibrotic factors. In this study, we investigated the correlation of the mRNA expression of podocyte proteins and profibrotic factors with renal fibrosis measured in renal biopsies of patients with primary and secondary glomerulopathies. METHODS: Eighty-four adult patients with primary or secondary glomerular diseases and 12 controls were included. Demographic and clinical data were collected. Seventy-two percent of the renal biopsies were done less than one year from clinical disease manifestation. The quantification of the podocyte-associated mRNAs of alpha-actinin-4, podocin, and podocalyxin, as well as of the profibrotic factors TGF-ß1, CTGF, and VEGF-A were quantified by real-time polymerase chain reaction. The percent positive area of renal fibrosis was measured by immunohistochemistry staining, using anti-CTGF and anti-HHF35 antibodies and unpolarized Sirius Red. Correlations between the expression of tissue mRNAs and the positive area of fibrosis for the measured markers were made by Spearman's rank correlation coefficient. RESULTS: In relation to control biopsies, podocyte-specific proteins were downregulated in podocytopathies, in proliferative nephritis, in diabetic kidney disease (DRD), and in IgA nephropathy (IgAN). Messenger RNA of TGF-ß1, CTGF, and VEGF-A was upregulated in patients with podocytopathies and in DRD but not in proliferative nephritis and IgAN. Tissue mRNA expression of TGF-ß1, CTGF, and VEGF-A were strongly correlated with renal fibrosis, as measured by HHF35; however, the correlation, albeit significant, was moderate for Sirius Red and weak for CTGF. The percent positive area of renal fibrosis measured by Sirius Red was similar between podocytopathies and DRD and significantly higher in podocytopathies compared to IgAN or proliferative nephritis. CONCLUSIONS: In patients with glomerular diseases, the mRNA of TGF-ß1, CTGF, and VEGF-A correlated positively with the extent of renal fibrosis, and the positive area of fibrosis was larger in the podocytopathies and in DRD as measured by Sirius Red. The pathways connecting podocyte damage and activation of profibrotic factors to kidney tissue fibrosis need to be better investigated.
Assuntos
Glomérulos Renais/patologia , Podócitos/patologia , Adulto , Biomarcadores/metabolismo , Biópsia , Fator de Crescimento do Tecido Conjuntivo/genética , Feminino , Humanos , Glomérulos Renais/metabolismo , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , Fator de Crescimento Transformador beta1/genética , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
AIMS: Since lupus nephritis (LN) etiopathogenesis is not fully understood, herein we investigated the morphological basis of LN in mice induced with pristane. MAIN METHODS: To evaluate the melatonin effects in these animals, we studied the renal cytoarchitecture by means of morphological analyses, immunofluorescence expression of specific markers related to fibrosis, oxidative stress, inflammation and apoptosis. KEY FINDINGS: We observed that pristane-LN mice have serious alterations in the kidney cytoarchitecture, i.e. tubular degeneration, glomerular hypercellularity, matrix mesangial expansion and interstitial inflammation. The pristane-induced LN mice treated with melatonin exhibited a well preserved cytoarchitecture. SIGNIFICANCE: Our results document that LN etiopathogenesis is related to both tubular damage and glomerular lesions. We suggest that it is essential to take in consideration both these lesions for LN diagnosis and classification. Clearly, we show that the use of melatonin may be a possible therapeutic strategy for improvement the renal injury in this disorder.
Assuntos
Nefrite Lúpica/tratamento farmacológico , Melatonina/uso terapêutico , Animais , Apoptose , Autoanticorpos/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Fibrose , Inflamação/patologia , Rim/lesões , Rim/metabolismo , Rim/patologia , Nefropatias/patologia , Glomérulos Renais/metabolismo , Nefrite Lúpica/metabolismo , Nefrite Lúpica/prevenção & controle , Melatonina/metabolismo , Melatonina/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo , Substâncias Protetoras , TerpenosRESUMO
INTRODUCTION: Lupus nephritis (LN) is one of the most severe complications of systemic lupus erythematosus. As murine models of LN are valuable tools to better understand its pathophysiology and to search for new effective treatments, we investigated the effects of the bioflavonoid quercetin on pristane-induced LN mice through histomorphological analyses. METHODS: Immunofluorescence and biochemical assays were used to evaluate the expression of markers of inflammation (interleukin-6, IL-6; tumour necrosis factor-α, TNF-α), oxidative stress (catalase, CAT; superoxide dismutase 1, SOD1; thiobarbituric acid reactive substances, TBARS), apoptosis (Bax), and fibrosis (transforming growth factor-ß1, TGF-ß1). Glomerular and tubular ultrastructure was analysed, and tissue messenger RNA of podocin, podoplanin and α3ß1-integrin were quantified using the real-time polymerase chain reaction. RESULTS: Pristane-induced LN mice showed severe kidney injury, characterized by increased proteinuria, glomerular mesangial expansion and inflammation, high expression of the pro-fibrotic, apoptotic and prooxidant markers and reduction of antioxidants. In the kidney ultrastructure, foot process (FP) effacement, apoptotic mesangial cells and abnormal mitochondria with disrupted cristae were observed, along with suppressed tissue mRNA of podocin, podoplanin and α3ß1-integrin. Treatment with quercetin in the pristane-induced LN mice model was nephroprotective, decreasing proteinuria levels and significantly lowering tissue expression of IL-6, TNF-α, TGF-ß1, Bax and TBARS. Simultaneously, quercetin significantly increased CAT and SOD1 expressions in these mice. In addition, it was observed improvement of the kidney ultrastructure, and tissue mRNA of podocin, but not podoplanin and α3ß1-integrin, was restored to the levels found in the control mice. CONCLUSION: In conclusion, these findings provide experimental evidence of the renoprotective effects of quercetin in the pristane-induced LN mice model. We suggest that quercetin effectively ameliorates the kidney damage caused by pristane, a bioflavonoid to be further evaluated as a new therapeutic strategy in this disease.
Assuntos
Injúria Renal Aguda/tratamento farmacológico , Antioxidantes/uso terapêutico , Glomérulos Renais/patologia , Nefrite Lúpica/tratamento farmacológico , Quercetina/uso terapêutico , Injúria Renal Aguda/patologia , Injúria Renal Aguda/prevenção & controle , Animais , Catalase/biossíntese , Citocinas/biossíntese , Modelos Animais de Doenças , Feminino , Inflamação/patologia , Nefrite Lúpica/induzido quimicamente , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Proteinúria/tratamento farmacológico , Superóxido Dismutase-1/biossíntese , TerpenosRESUMO
OBJECTIVES: Systemic lupus erythematosus (SLE) is an autoimmune inflammatory disease with renal involvement in over half of the cases. In lupus nephritis (LN), podocytes are injured at the structural and molecular level. Spontaneous or induced animal models of SLE can reproduce the glomerular damage, similar to what is observed in humans. In this review, murine models focusing the podocyte injury were summarized, and therapeutic strategies to protect the podocyte cell were explored. METHODS: Using the PubMed and MEDLINE databases from 1950 to 2015, literature search was conducted by article title and abstract, combining the following key words: "systemic lupus erythematosus," "lupus nephritis," "animal model," "podocyte injury," and "treatment." RESULTS: Published or in-press eligible studies that were published as full-length articles in English-language journals were considered. Articles were summarized according to podocyte structure and function, the podocyte injury resulting from spontaneous (NZB/W F1 hybrid, MRL/lpr, and BXSB-Yaa mice) or induced (chronic graft-versus-host disease and pristane) mice models of LN, and the protective effects of drug treatments on podocyte cell structure and function reported in these models. CONCLUSIONS: Murine models of SLE have proven useful for better comprehension of the multiple mechanisms involved in systemic autoimmunity that leads to LN. These critical tools should be considered when target therapies are designed to control this disorder.
Assuntos
Imunossupressores/administração & dosagem , Nefrite Lúpica/tratamento farmacológico , Podócitos/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Humanos , Imunossupressores/farmacologia , Nefrite Lúpica/patologia , Camundongos , Camundongos Endogâmicos MRL lpr , Camundongos Endogâmicos NZB , Podócitos/patologiaRESUMO
OBJECTIVE: The aim of this study was to correlate different degrees of excess weight with the expression of podocyte-associated messenger RNAs (mRNAs) in urine. METHODS: The sample comprised 83 patients with overweight or obesity class I, II, or III and 18 healthy controls. The expression of nephrin, podocin, podocalyxin, α-actinin-4, α3ß1integrin, vascular endothelial growth factor, and transforming growth factor-beta (TGF-ß1 ) mRNA in urine was quantified with the real-time polymerase chain reaction. mRNA expression was correlated with body mass index, the metabolic syndrome, albuminuria, and inflammation. RESULTS: Adults with obesity class III had higher levels of serum lipids, glucose, HbA1C, insulin resistance, and C-reactive protein (P < 0.05), with 85% of the subjects meeting criteria for the metabolic syndrome (P < 0.001 vs. other groups). Urinary podocyte-associated mRNAs were higher in adults with obesity class III than in other groups (P < 0.05). Patients with overweight or obesity class I or II also had higher levels of podocyte mRNAs than controls: nephrin (P = 0.021), α-actinin-4 (P = 0.014), α3ß1integrin (P = 0.036), and TGF-ß1 (P = 0.005). Metabolic syndrome, hyperinsulinemia, and C-reactive protein were correlated with podocyturia, but only higher insulin levels were related regardless of obesity. CONCLUSIONS: Severe obesity and hyperinsulinemia were associated with higher urinary expression of podocyte-associated mRNAs, even at normal urinary albumin excretion rates.
Assuntos
Obesidade Mórbida/urina , Podócitos/metabolismo , RNA Mensageiro/metabolismo , RNA Mensageiro/urina , Adulto , Estudos Transversais , Feminino , Humanos , Hiperinsulinismo , Masculino , Pessoa de Meia-Idade , Obesidade Mórbida/metabolismo , Podócitos/química , RNA Mensageiro/químicaRESUMO
AIM: Glomerular deposition of immune complexes and inflammation induce podocyte injury in lupus nephritis (LN). This study hypothesized that the severity of the histological lesions of LN affects podocyte-associated mRNAs profiles in kidney tissue and in urine. METHODS: Thirty-three patients with LN were grouped according to the presence of mild mesangial (classes I and II) or moderate-to-severe immune complex deposition, proliferation and/or inflammation (classes III, IV and V) in kidney biopsy. Tissue and urine mRNA of nephrin, podocin, podocalyxin, α-actinin-4, transient receptor potential cation channel 6, and of growth factors VEGF-A and TGF-ß1 and the transcription factor FOXP3 were measured using real time polymerase chain reaction. These mRNAs were correlated with histological severity of LN, extent of glomerular immune deposits, and tissue infiltrating cells. RESULTS: Podocyte-associated mRNAs were inhibited in renal tissue of patients with LN irrespective of histological class when compared to controls. However, significantly higher expression of podocyte mRNAs in urine, including those of growth factors and FOXP3, were found in patients with moderate-to-severe nephritis, mostly in class III and IV proliferative forms. The number of invading CD8+ T cells, B cells and macrophages correlated positively with urine podocyte-associated mRNAs. Urine podocyte mRNAs also correlated with proteinuria. CONCLUSIONS: Inhibition of podocyte-associated mRNAs in kidney tissue suggests that podocyte injury occurs regardless of class severity of LN. Increased urinary excretion of podocyte mRNAs, mostly in patients with moderate-to-severe lesions, may reflect a greater burden of glomerular damage with detachment of podocytes into the urine.
Assuntos
Perfilação da Expressão Gênica , Nefrite Lúpica/genética , Podócitos/química , RNA Mensageiro/genética , Adolescente , Adulto , Idoso , Biópsia , Estudos de Casos e Controles , Feminino , Perfilação da Expressão Gênica/métodos , Marcadores Genéticos , Humanos , Nefrite Lúpica/diagnóstico , Nefrite Lúpica/imunologia , Nefrite Lúpica/urina , Masculino , Pessoa de Meia-Idade , Podócitos/imunologia , Podócitos/patologia , Valor Preditivo dos Testes , Prognóstico , RNA Mensageiro/urina , Reação em Cadeia da Polimerase em Tempo Real , Índice de Gravidade de Doença , Urinálise , Urina/citologia , Adulto JovemRESUMO
AIM: It is not clear how the podocyte damage manifests in different glomerulopathies. This study evaluated the podocyte-associated mRNA profiles in renal tissue and urine of patients with proliferative (PGs) or non-proliferative (NPGs) glomerulopathies. METHODS: Messenger RNA levels of nephrin, podocin, podocalyxin, synaptopodin, and alpha-actinin-4 were measured in the kidney tissue and urinary cells by real-time polymerase chain reaction. Podocyte-associated mRNAs were correlated with proteinuria and renal function, and the effect of immunosuppressive treatment of PGs and NPGs on urine mRNAs was assessed up to one year of follow up. RESULTS: Podocyte-associated mRNAs were expressed consistently less in kidney tissue from patients with NPGs, and urinary podocyte mRNA levels were significantly higher in the PG group. After six months of immunosuppressive therapy, patients with PGs showed a significant reduction in the expression of podocin, podocalyxin, and alpha-actinin-4 compared with baseline (p<0.001). In the NPG group, alpha-actinin-4 levels decreased (p=0.008), and there was also a trend toward reduced podocalyxin mRNA (p=0.08). Urine podocyte-associated mRNAs correlated with the level of proteinuria at baseline and at six months, and there was a trend toward an inverse correlation between urinary mRNAs and kidney function at one year of follow up. CONCLUSIONS: Podocyte-associated mRNAs were inhibited in kidney tissue concomitantly with their increase in urine in these patients with glomerulopathies. Different profiles of mRNA expression were seen, pointing to a higher degree of intra-renal podocytopenia in the NPGs and of podocyturia in the PGs. The immunosuppressive therapy effectively reduced the urinary levels of podocyte-associated mRNAs.